Ex vivo analysis of human antigen-specific CD8+ T-cell responses:: Quality assessment of fluorescent HLA-A2 multimer and interferon-γ ELISPOT assays for patient immune monitoring
Ex vivo analysis of human antigen-specific CD8+ T-cell responses:: Quality assessment of fluorescent HLA-A2 multimer and interferon-γ ELISPOT assays for patient immune monitoring
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DOI：
10.1097/00002371-200407000-00006
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发表时间：
2004-07-01
影响因子：
3.9
通讯作者：
Romero, P
中科院分区：
文献类型：
--
作者：
Speiser, DE;Pittet, MJ;Romero, P
作者开发了一种用于免疫监测外周血淋巴细胞（PBL）内抗原特异性CD 8 + T细胞的标准化方法，该方法将Melan-A/MART-1和流感特异性CD 8 + T细胞的直接离体分析与HLA-A2/肽多聚体和干扰素-γ ELISPOT测定相结合。在这里，作者评估了从健康供体和黑色素瘤患者中获得的180个PBL结果的质量。多聚体测定的重现性良好（平均15%变化）。在缺乏体内抗原特异性T细胞应答的情况下，多聚体阳性T细胞的生理波动较低，Melan-A的变异系数为20%，流感特异性T细胞的变异系数为28%。相比之下，具有疫苗诱导的T细胞应答的患者的T细胞频率显著增加，明显超过生理波动。用ELISPOT测定获得了相当的结果。总之，这种方法非常适合评估T细胞应答作为临床疫苗研究中的生物学终点。
The authors developed a standardized approach for immune monitoring of antigen-specific CD8+ T cells within peripheral blood lymphocytes (PBLs) that combines direct ex vivo analysis of Melan-A/MART-1 and influenza-specific CD8+ T cells with HLA-A2/peptide multimers and interferon-gamma ELISPOT assays. Here the authors assessed the quality of results obtained with 180 PBLs from healthy donors and melanoma patients. Reproducibility of the multimer assay was good (average of 15% variation). In the absence of in vivo antigen-specific T-cell responses, physiologic fluctuations of multimer-positive T cells was low, with variation coefficients of 20% for Melan-A and 28% for influenza-specific T cells. In contrast, patients with vaccination-induced T-cell responses had significantly increased T-cell frequencies clearly exceeding physiologic fluctuations. Comparable results were obtained with ELISPOT assays. In conclusion, this approach is well suited to assess T-cell responses as biologic endpoints in clinical vaccine studies.